Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, 9177948974 Mashhad, Iran.
Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, 9177948974 Mashhad, Iran.
Life Sci. 2021 Jun 1;274:119336. doi: 10.1016/j.lfs.2021.119336. Epub 2021 Mar 11.
This study aimed to explore the therapeutic effects of amniotic fluid-derived extracellular vesicles including exosomes (AF-Exos) on the recovery of sperm production capacity in a rat model of azoospermia.
The non-obstructive azoospermia (NOA) was induced in rats using intratesticular administration of Busulfan. Azoospermia was confirmed by testis histology. AF-Exos samples containing 10 or 40 μg exosomal proteins were injected into testicular tissue of NOA rats. After two months, the recovery of spermatogenesis was monitored via histopathological staining, spermiogram, and hormonal analysis. Immunohistochemistry staining for OCT-3/4 was used to identify of spermatogonial progenitors. The expression of DAZL and VASA, was also measured.
AF-Exos exhibited sphere-shaped morphology with the mean diameter and zeta potential of 50 ± 7.521 nm and -7.16 mV. Immunoblots revealed that isolated nanoparticles were CD63, CD9, and CD81 positive. Histopathological evaluation revealed that spermatogenesis was improved significantly in NOA rats after AF-Exos injection. Data showed that the sperm parameters and spermatogenesis index were significantly improved after AF-Exos injection compared to azoospermic groups. OCT-3/4 cells were increased in NOA rats after AF-Exos injection, showing the restoration of spermatogenesis. In the present study, both doses of exosome (10 and 40 μg) restored the testicular function of NOA rats. DAZL and VASA were increased significantly in animals who received 40 μg exosomal protein compared to azoospermic rats. Except in a high dose of AF-Exos (40 μg) for Testosterone and FSH, no statistically significant differences were found regarding hormones post-exosome injection.
Our study demonstrated that AF-Exos regenerated spermatogenesis and improved sperm quality in NOA rats.
本研究旨在探讨羊水衍生的细胞外囊泡（包括外泌体）对布硫苯脲诱导的大鼠无精子症模型中精子生成能力恢复的治疗作用。
通过睾丸内给予布硫苯脲诱导非阻塞性无精子症（NOA）大鼠，通过睾丸组织学确认无精子症。将含有 10 或 40μg 外泌体蛋白的 AF-Exos 样本注射到 NOA 大鼠的睾丸组织中。两个月后，通过组织病理学染色、精子计数和激素分析监测生精恢复情况。免疫组化染色用于鉴定精原细胞祖细胞的 OCT-3/4。还测量了 DAZL 和 VASA 的表达。
AF-Exos 呈球形形态，平均直径和 Zeta 电位分别为 50±7.521nm 和-7.16mV。免疫印迹显示分离的纳米颗粒呈 CD63、CD9 和 CD81 阳性。组织病理学评估显示，AF-Exos 注射后 NOA 大鼠的生精明显改善。与无精子症组相比，AF-Exos 注射后精子参数和生精指数显著提高。AF-Exos 注射后 NOA 大鼠的 OCT-3/4 细胞增加，表明生精恢复。在本研究中，两种剂量的外泌体（10 和 40μg）均恢复了 NOA 大鼠的睾丸功能。与无精子症大鼠相比，接受 40μg 外泌体蛋白的动物的 DAZL 和 VASA 显著增加。除了高剂量的 AF-Exos（40μg）对睾酮和 FSH 外，注射外泌体后激素没有统计学上的显著差异。
我们的研究表明，AF-Exos 可再生精子发生并改善 NOA 大鼠的精子质量。
